Cat No.: 31109 Other Names: P1NP, N-Propeptide of Type 1 Procollagen, PINP
Introduction
Procollagen type 1 N-terminal propeptide (P1NP) is the N-terminal extension of procollagen type 1 removed proteolytically during
collagen formation1. P1NP is a bone formation biomarker usually secreted into blood stream as unstable trimeric structure which
subsequently undergo thermal degradation to form a stable monomeric structure2.
Principle of the Assay
This assay is a quantitative sandwich ELISA. The immunoplate is pre-coated with a monoclonal antibody specific for human P1NP.
Standards and samples are pipetted into the wells and any human P1NP present is bound by the immobilized antibody. After
washing away any unbound substances, a biotin labelled polyclonal antibody specific for human P1NP is added to the wells. After
wash step to remove any unbound reagents, streptavidin-HRP conjugate (STP-HRP) is added. After the last wash step, an HRP
substrate solution is added and colour develops in proportion to the amount of human P1NP bound initially. The assay is stopped
and the optical density of the wells determined using a microplate reader. Since the increases in absorbance are directly proportional to the amount of captured human P1NP, the unknown sample concentration can be interpolated from a reference curve included in
each assay.
Assay Performance
A. Typical representation of standard curve
The following standard curve is provided for demonstration only. A standard curve should be generated for each set of sample
assay.
Human P1NP (pg/ml) | Absorbance (450 nm) | Blanked Absorbance |
0 | 0.094 | 0 |
62.5 | 0.133 | 0.039 |
125 | 0.187 | 0.093 |
250 | 0.293 | 0.199 |
500 | 0.474 | 0.38 |
1000 | 0.812 | 0.718 |
2000 | 1.317 | 1.223 |
4000 | 2.096 | 2.002 |
B. Sensitivity
The lowest level of human PINP that can be detected by this assay is 62.5 pg/mL.
C. Precision
Intra-assay Precision (Precision within an assay) C.V. <3.3%.
Inter-assay Precision (Precision between assays) C.V.<5.1%.